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. 2020 Mar 31;9:e49392. doi: 10.7554/eLife.49392

Figure 2. GPC1-specific human hCAR-T cells specifically recognized hGPC1-positive tumor cells and inhibited tumor growth in xenograft mouse model.

(A) Diagrams of GPC1-specific human hCAR; scFv frgments derived from light chain (VL) and heavy chain (VH) of anti-GPC1 mAb (clone: 1–12) were fused to human CD28 and human CD3ζ signal domains. The positions of VL and VH were switched to generate two forms of CAR gene, LH and HL. (B) LK2-hGPC1, LK2-mock, and endogenous hGPC1-expressing TE14 were stained by anti-GPC1 mAb (clone: 1–12) (shaded histogram) or isotype control (open histogram). (C) GPC1-specific IFNγ secretion of hCAR-T cells (LH or HL form) or hCont-T cells co-cultured with LK2-mock, LK2-hGPC1, or TE14. (D) Antigen-specific in vitro cytotoxicity of hCAR-T cells (LH or HL form) or hCont-T cells against LK2-hGPC1, LK2-mock, or TE14 was evaluated by using standard Cr51 releasing assay. (E) hCAR-T cells (LH or HL form) or hCont-T cells (2 × 107 cells/mouse) were injected into TE14-bearing NOG mice on day 9. Results are representative of two or three experiments. Error bars indicate SD.

Figure 2.

Figure 2—figure supplement 1. The sequences of GPC1-specific human CAR vectors.

Figure 2—figure supplement 1.

The sequences of the hCAR vectors (LH and HL forms) are shown. The hCAR vectors are comprised of the human CD8a leader sequence (red), scFv of the anti-GPC1 mAb (linker; orange and VL; purple or VH; green), human CD28 extracellular/transmembrane/intracellular domains (light blue), and human CD3ζ intracellular domain (dark blue).
Figure 2—figure supplement 1—source data 1. The sequences of GPC1-specific human CAR vectors.